TNF-α、Caspase-3在大鼠脊髓损伤后神经细胞凋亡中相关性研究  

The correlation of TNF-α and Caspase-3 in nerve cell apoptosis after spinal cord injury of rats

作　　者：刘铭[1] 刘欣伟[1] 刘宪民[1] 刘松波[1] 王宇[1] 

机构地区：[1]沈阳军区总医院骨科全军重症战创伤救治中心,辽宁沈阳110016

出　　处：《临床军医杂志》2017年第4期366-370,共5页Clinical Journal of Medical Officers

基　　金：辽宁省自然科学基金资助项目(2013020175);辽宁省科技攻关项目(2013225089)

摘　　要：目的探讨大鼠脊髓损伤后肿瘤坏死因子-α(TNF-α)、天冬氨酸特异性半胱氨酸蛋白酶-3(Caspase-3)在神经细胞凋亡中的动态表达,验证TNF-α和Caspase-3之间的相关性。方法将70只成年健康SD大鼠按Nystrom法建立大鼠脊髓(T8、T9)急性压迫损伤模型,HE染色观察脊髓组织病理学变化;免疫组化染色测定各时间点TNF-α、Caspase-3的表达变化;原位末端脱氧核糖核酸转移酶介导的脱氧尿苷三磷酸(d UTP)标记法(TUNEL法)检测神经细胞的凋亡水平;通过Western blot法检测大鼠脊髓中TNF-α、Caspase-3蛋白的表达水平。结果正常大鼠脊髓神经细胞TNF-α、Caspase-3的阳性细胞表达率较少;脊髓损伤后8 h,脊髓神经细胞中TNF-α、Caspase-3的阳性表达率明显增多,分别为(24.13%±3.02%)及(40.79%±3.43%),1 d达高峰,分别为(51.36%±4.26%)及(70.78%±5.97%),3 d表达减弱,分别为(20.24%±2.93%)及(37.71%±2.88%),与正常大鼠比较,差异有统计学意义(P<0.01)。TUNEL阳性细胞率在大鼠脊髓损伤后8 h明显增多(39.81%±2.27%),1 d达高峰(71.58%±3.87%),3 d表达减弱(40.55%±2.36%),与正常大鼠比较,差异均有统计学意义(P<0.01)。大鼠脊髓损伤后1 d,与损伤组比较,抑制剂组TNF-α、Caspase-3、TUNEL阳性细胞率明显减少,差异有统计学意义(P<0.01)。造模后7 d,与空白对照组比较,损伤组TNF-α、Caspase-3蛋白的表达上升;与损伤组比较,抑制剂组TNF-α、Caspase-3蛋白的表达下降,差异有统计学意义(P<0.05)。结论脊髓损伤后TNF-α、Caspase-3的表达增强,TNF-α抑制剂使Caspase-3表达减弱,且其参与了脊髓损伤细胞凋亡的调节。Objective To investigate dynamic expression in the nerve cell apoptosis, and verify the correlation between TNF-α and Caspase-3. Methods The spinal cord injury of 70 healthy adult SD rats were induced in Nystrom way by the moderate compression at the level of T8 and T9 spinal cord. HE methods were used to detect the pathologic change of spinal cord. The expression of TNF-α and Caspase-3 were detected by immunohistochemical study. Besides, using the terminal deoxynucleotidyl transferase mediated DUTP nick and labeling（TUNEL）methods to detect the level of the apoptosis of neural cells. Western-blot to detect laboratory animals TNF-α Caspase-3 protein expression level. Results The positive ceils expression rate of TNF-α and Caspase-3 in normal rat spinal cord neurons was less;after 8 hours of spinal cord injury, the positive cells expression rate of TNF-α and Caspase-3 in normal rat spinal cord neurons were significantly increased at（24. 13% ± 3.02% ）and （40.79 % ± 3.43% ）, retrospectively. One day later reached the peak, （51.36% ± 4. 26% ） and （ 70. 78% ± 5.97% ）, retrospectively. Three days later began to reduce to （ 20. 24% ± 2. 93% ） and （37.71% ± 2. 88 % ）, retrospectively. Compared with normal rats, the differences were statistically significant （P 〈 0. 01 ）. The positive cell expression rate of TUNEL significantly increased after 8 hours of spinal cord injury （39. 81% ± 2. 27% ）. One day later reached the peak（71.58% ±3.87% ）. Three days later began to reduce at （40. 55% ± 2. 36% ）. Compared with the normal rats, and the differences were statistically significant （P 〈 0. 01 ）. One day after spinal cord injury, compared with the injury group, the positive cells rate of TNF-α, Caspase-3 and TUNEL in inhibitor group significantly reduced, and the difference had statistical significance （ P 〈 0. 01 ）. Seven days after modeling,compared with the control group,the expression level of TNF-α and Caspase-3 in the injury group increased;compared

关 键 词：脊髓损伤 细胞凋亡 肿瘤坏死因子-Α 天冬氨酸特异性半胱氨酸蛋白酶-3 

分 类 号：R285.5[医药卫生—中药学]